| Literature DB >> 36173567 |
Ryoya Hayashi1,2, Azusa Inoue3,4.
Abstract
Cleavage Under Target & Release Using Nuclease (CUT&RUN) enables the detection of DNA regions that are bound by a protein of interest. This method is suitable for low-input materials because of the absence of an immunoprecipitation step. However, it sometimes fails when applying it to fragile cells, such as mouse oocytes. Here we describe our low-input CUT&RUN protocol optimized for mouse oocyte and preimplantation embryo samples in which the primary antibody and protein A-MNase binding steps are completed before the cells are bound to Concanavalin A-coated magnetic beads. This modification prevents crush of oocytes and early embryos and unwanted loss of chromatin during CUT&RUN procedures.Entities:
Keywords: Epigenetics; Histone modification; Oocytes; Preimplantation embryo
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Year: 2023 PMID: 36173567 DOI: 10.1007/978-1-0716-2724-2_6
Source DB: PubMed Journal: Methods Mol Biol ISSN: 1064-3745